Fig. 3: Quantitative Uptake of extracellular mitochondria.

A Unlabeled HeLa acceptor cells were incubated for 24 hours with fractions F4, F4′, or F16 derived from donor cells expressing Nluc-HA-OMP25 or COX8a-Nluc-HA. NLuc activity was measured after 24 h. Input NLuc activity was set to 100%. Three independent experiments were performed, each including three technical replicates. Each dot represents a replicate. Error bars indicate standard deviation, center represent the means. *Statistical significance was assessed by unpaired, two-tailed t-test comparing F4 or F4′ to F16; p < 0.01. B, C Uptake kinetics and dose response. Acceptor cells were incubated with F4 or F4′ (from Nluc-HA-OMP25-expressing cells, 50,000 total NLuc activity units) for various durations B, or for 24 h with increasing amounts of F4 or F4′ input C. Three independent experiments were performed, each including four technical replicates. Each dot represents a replicate. D Temperature block experiment. Unlabeled acceptor cells were incubated with F4 and F4′ (from Nluc-HA-OMP25-expressing cells) for 4 h at either 37 °C or 4 °C. NLuc activity was then measured. Three independent experiments were performed, each including four technical replicates. Each dot represents a replicate. Statistical significance was assessed by unpaired, two-tailed t-test comparing F4 or F4′ at 37 °C vs. 4 °C; p < 0.01.